Anti-angiogenic agents and anti-obesity substances applied with anti-angiogenesis from natural products

ABSTRACT

Disclosed herein are an anti-obesity agent from natural products based on angiogenesis inhibitory actions, and an angiogenesis inhibitor or a composition for inhibiting obesity, containing at least one selected from the group consisting of  Psoraleae  Semen extract,  Siegesbeckiae  Herba extract, and Corni Fructus extract. Because the  Psoraleae  Semen extract,  Siegesbeckiae  Herba extract, and Corni Fructus extract of the present invention exhibit angiogenesis inhibitory effects and obesity inhibitory effects, they may be usefully used as an agent for prevention and treatment of angiogenesis-related diseases or obesity.

CROSS-REFERENCES TO RELATED APPLICATION

This patent application claims the benefit of priority from KoreanPatent Application No. 10-2009-0027470, filed on Mar. 31, 2009, thecontents of which are incorporated herein by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present disclosure relates to an anti-obesity substance applied withanti-angiogenesis from natural products.

2. Description of the Related Art

The term “angiogenesis” refers to a series of processes involving thegeneration of new blood vessels from pre-existing blood vessels, and isknown to be essentially required in normal anatomical functions such asfetal development, female menstruation, wound healing, etc.; and variousdiseases such as growth of cancer and mast cells, rheumatoid arthritis,diabetic blindness, etc. However, it is a very rigorously controlledphenomenon that rarely occurs under normal conditions. In an organism,angiogenesis inducing factors maintain an equilibrium state withangiogenesis inhibiting factors under normal conditions. However,because the number of vascular growth promoting factors increases orvascular growth inhibiting factors do not function properly underdisease-causing circumstances, angiogenesis may not be autonomouslyregulated and may continue to grow, leading to development of disease.

The formation of angiogenesis has been known to be promoted by 20 ormore vascular formation promoting factors until now, and vascularendothelial growth factor (VEGF) among them is secreted in various kindsof tumor and mast cells and is known to be the most potent vascularformation promoting factor. The VEGF is also known as a vascularpermeability factor and known to bind to the receptors thereof, VEGFR-1(Flt-1) and VEGFR-2 (Flk-1/KDR), to cause the proliferation of vascularendothelial cells and increase vascular permeability to be involved inthe growth and metastasis of tumor and mast cells (Leung D W et al.,Science 246: 1306-1309, 1989; Liping Liu & Meydani Mohsen NutritionReview, 61(11): 384-387, 2003; Jaap G et al., The FASEB Journal10.1096/fj.03-1101fje. 2004; Hausman G J & Tichardson R L, J. anim.Sci., 82: 925-934, 2004).

In particular, angiogenesis plays an important role in formation of mastcells and rapidly supplies oxygen and nutrients to obesity tissues todirectly aid in the growth of obesity tissues. Therefore, the formationof a new blood vessel in any tissue is inhibited by inhibiting theformation of the new blood vessel to prevent the tissue fromhypertrophy. Folkman (PNAS 99: 10730-10735, 2002) reported that a mouselost weight when an angiogenesis inhibitor was administered, andNishimura et al., reported that the formation of a new blood vesseldirectly promoted the growth of mast cells.

Obesity may be defined as a state in which body fat is excessivelyaccumulated to harm the health, and is established as a recognized‘chronic disease’ which must be treated because it was revealed that itis responsible for social and psychological disorder as well asincreases to mortality and morbidity due to various chronic degenerativediseases such as hypertension, arteriosclerosis, coronary heart disease,type 2 diabetes mellitus, fatty liver, hyperlipidemia, degenerativearthritis, some cancer diseases, etc. Current obesity drugs may includefenfluramine and sibutamine preparations that are suppressors ofappetite, which reduce food uptake, and ephedrine and orlistatpreparations that inhibit lipogenesis or increase metabolic activity.However, these drugs cause many side effects (alopecia) and addictionproblems emerge. Therefore, there is need for development of ananti-obesity agent from natural products.

Thus, the present inventors have discovered Psoraleae Semen extract,Siegesbeckiae Herba extract, and Corni Fructus extract, which exhibitinhibiting effects of angiogenesis among various extracts from naturalproducts, found that these exhibit anti-obesity effects, confirmed thatPsoraleae Semen extract, Siegesbeckiae Herba extract, and Corni Fructusextract of the present invention may be used as active ingredients foran angiogenesis inhibitor or anti-obesity composition, thereby leadingto completion of the present invention.

SUMMARY OF THE INVENTION

One object of the present invention is to provide an angiogenesisinhibitor, a composition for prevention or treatment ofangiogenesis-related diseases, or a composition for anti-obesity withthe inhibitory activity of angiogenesis, containing Psoraleae Semenextract, Siegesbeckiae Herba extract, and Corni Fructus extract asactive ingredients.

Another object of the present invention is to provide a method forinhibiting angiogenesis, a method for preventing or treatingangiogenesis-related diseases, or a method for inhibiting obesity,including administering to an individual an effective amount ofPsoraleae Semen extract, Siegesbeckiae Herba extract, and Corni Fructusextract.

In order to achieve the objects, the present invention provides anangiogenesis inhibitor containing at least one selected from the groupconsisting of Psoraleae Semen extract, Siegesbeckiae Herba extract, andCorni Fructus extract as active ingredients.

The present invention also provides a method for inhibitingangiogenesis, including administering to an individual at least oneselected from the group consisting of Psoraleae Semen extract,Siegesbeckiae Herba extract, and Corni Fructus extract.

Furthermore, the present invention provides a composition for preventionor treatment of angiogenesis-related diseases, containing at least oneselected from the group consisting of Psoraleae Semen extract,Siegesbeckiae Herba extract, and Corni Fructus extract as an activeingredient.

The present invention also provides a method for preventing or treatingangiogenesis-related diseases, including administering to an individualat least one selected from the group consisting of Psoraleae Semenextract, Siegesbeckiae Herba extract, and Corni Fructus extract.

Furthermore, the present invention provides a composition foranti-obesity with the inhibitory activity of angiogenesis, containingPsoraleae Semen extract or Siegesbeckiae Herba extract as activeingredients.

The present invention also provides a method for inhibiting obesity,including administering to an individual an effective amount ofPsoraleae Semen extract or Siegesbeckiae Herba extract.

Furthermore, the present invention provides a health supplement food forpreventing or improving antiogenesis-related diseases such ascerebrovascular diseases, cardiovascular diseases, ocular diseases orcancerous diseases, containing at least one selected from the groupconsisting of Psoraleae Semen extract, Siegesbeckiae Herba extract, andCorni Fructus extract as an active ingredient.

In addition, the present invention provides a health supplement food forpreventing or improving obesity, containing Psoraleae Semen extract orSiegesbeckiae Herba extract as an active ingredient.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawings will be provided by the Office upon request and paymentof the necessary fee.

The above and other objects, features and other advantages of thepresent invention will be more clearly understood from the followingdetailed description taken in conjunction with the accompanyingdrawings, in which:

FIG. 1 is a group of graphs confirming the cytotoxicity of each extractof Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructus.

FIG. 2 is a group of graphs illustrating inhibitory effects of eachextract of Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructus on acell adhesion promoting factor VACM-1.

FIG. 3 is a group of graphs illustrating inhibitory effects of eachextract of Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructus on acell adhesion promoting factor IACM-1.

FIG. 4 is a group of graphs illustrating inhibitory effects of eachextract of Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructus on acell adhesion promoting factor E-selectin.

FIG. 5 is a graph illustrating angiogenesis inhibitory effects by eachextract of Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructus.

FIGS. 6 through 8 are a group of photos illustratingexpression-inhibitory effects of each extract of Psoraleae Semen,Siegesbeckiae Herba, and Corni Fructus on signal transduction molecules:

FIG. 6: Psoraleae Semen;

FIG. 7: Siegesbeckiae Herba; and

FIG. 8: Corni Fructus.

FIG. 9 is a view illustrating the signal transduction pathway of asignal transduction molecule into an NF-kB.

FIG. 10 is a group of photos illustrating in vivo inhibitory effects ofeach extract of Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructusof the present invention on angiogenesis.

FIGS. 11 through 13 are groups of photos illustrating inhibitory effectsof each extract of Psoraleae Semen, Siegesbeckiae Herba, and CorniFructus of the present invention on adipocytes:

FIG. 11: Psoraleae Semen;

FIG. 12: Siegesbeckiae Herba; and

FIG. 13: Corni Fructus.

FIG. 14 is a view illustrating the control of lipogenesis andadipogenesis by signal transduction molecules SREBP-1 and PPARγ.

FIG. 15 is a view illustrating the inhibition of the adipocytedifferentiation resulting from decrease in the expression of PPARγ byeach extract of Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructusof the present invention.

FIG. 16 is a group of photos illustrating the inhibition of thelipogenesis and adipocyte differentiation resulting from decrease in theexpression of SREBP-1 and PPARγ by each extract of Psoraleae Semen,Siegesbeckiae Herba, and Corni Fructus of the present invention after acomplete adipocyte differentiation.

FIG. 17 is a graph illustrating the weight loss effects by each extractof Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructus of thepresent invention.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Features and advantages of the present invention will be more clearlyunderstood by the following detailed description of the presentpreferred embodiments by reference to the accompanying drawings. It isfirst noted that terms or words used herein should be construed asmeanings or concepts corresponding with the technical sprit of thepresent invention, based on the principle that the inventor canappropriately define the concepts of the terms to best describe his owninvention. Also, it should be understood that detailed descriptions ofwell-known functions and structures related to the present inventionwill be omitted so as not to unnecessarily obscure the important pointof the present invention.

Hereinafter, terms as used herein will be defined.

The term “prevention” as used herein means all the actions aimed atinhibiting the symptoms of angiogenesis-related diseases or obesity ordelaying the progression by administering a composition of the presentinvention.

The term “treatment” as used herein means all the actions aimed atalleviating or improving the symptoms of angiogenesis-related diseasesor obesity by administering a composition of the present invention.

The term “administration” as used herein means the introduction of adesired composition of the present invention into an individual in anyappropriate way.

The term “individual” as used herein means all the animals which have anangiogenesis-related disease or a disease in which the symptoms ofobesity may be alleviated, such as human, monkey, dog, goat, swine ormouse by administering a composition of the present invention to theanimals.

The term “pharmaceutically effective amount” as used herein means anamount sufficient to treat a disease at a reasonable benefit/dangerratio applied for a medical treatment, and this may be determineddepending on various factors known in the medical field, including thekind and severity of disease, drug activity, sensitivity, administrationtime, administration route and discharge ratio, administration timeperiod, co-administered drugs, and others.

Hereinafter, the present invention will be described in detail.

The present invention provides an angiogenesis inhibitor containing oneselected from the group consisting of Psoraleae Semen extract,Siegesbeckiae Herba extract, and Corni Fructus extract as an activeingredient.

The present invention also provides a method for inhibitingangiogenesis, including administering to an individual at least oneselected from the group consisting of Psoraleae Semen extract,Siegesbeckiae Herba extract, and Corni Fructus extract.

A Psoraleae Semen extract, Siegesbeckiae Herba extract, and CorniFructus extract of the present invention may be preferably prepared by apreparation method, including:

1) pulverizing each of dried Psoraleae Semen, Siegesbeckiae Herba, andCorni Fructus, followed by addition of an extraction solvent into eachfor extraction;

2) cooling down each of the extracts in Step 1, followed by filtration;and

3) evapoconcentrating each of the extracts in Step 2, followed byfreeze-drying, but is not limited thereto.

Whether cultivated or commercially available, the Psoraleae Semen,Siegesbeckiae Herba, and Corni Fructus may be used without anylimitation. The extracts of Psoraleae Semen, Siegesbeckiae Herba, andCorni Fructus according to the present invention may be prepared in thefollowing manner. Psoraleae Semen, Siegesbeckiae Herba, and CorniFructus are clearly washed with water, dried in the shade at roomtemperature and ground, placed into an extraction container, andextracted with an extraction solvent at an appropriate temperature for apredetermined time. A filter paper, etc. may be used to remove solidmatters from the obtained extracts, the suspension may be centrifuged,and then a supernatant may be filtered under vacuum. The extractionsolvent may be water, alcohol, or any mixture thereof, preferably oneselected from C₁₋₄ lower alcohol or any mixture thereof, and morepreferably ethanol, but is not limited thereto. The extraction solventis added in an amount of 2 to 10 times by dried weight of the PsoraleaeSemen, Siegesbeckiae Herba, and Corni Fructus. An extraction methodincluding hot water extraction, immersion extraction, reflux coldextraction, and ultrasonic extraction may be used, and preferably areflux cold extraction may be used 1 to 5 times. The extraction may beperformed preferably at 50° C. to 100° C., more preferably at 50° C. to60° C. The extraction may be performed for 1 to 7 hours, preferably for5 hours.

In the preparation method, the evapoconcentrating in Step 3) may beperformed preferably by an evaporator, and more preferably by rotaryvacuum evaporator, but is not limited thereto. The concentrating undervacuum may be performed preferably at 20° C. to 60° C., and morepreferably at 20° C. to 40° C., but is not limited thereto. Thefreeze-drying may be performed preferably by a freeze-dryer, morepreferably by a vibration freeze-dryer, but is not limited thereto. Thefreeze-drying may be performed preferably at −50° C. to −100° C., andmore preferably at −70° C., but is not limited thereto.

In a specific example of the present invention, the expression of celladhesion promoting factors is concentration-dependently inhibited in theorder of Psoraleae Semen>Siegesbeckiae Herba>Corni Fructus (See Tables 1to 3 and FIGS. 2 to 4) within the range (See FIG. 1) that each extractof Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructus does notexhibit the cytotoxicity. Each extract of the present inventionconcentration-dependently inhibited the expressions of signaltransduction molecules β-catenin and vascular epithelium-cadherin(VE-cadherin); and a subgroup Akt (Protein kinase B), and inhibited asignal transduction therefrom and blocked the signal transduction untilthe NF-kB (See FIG. 9), and exhibited inhibitory effects of angiogenesisformation, which were much better than those of ECC as a positivecontrol (See FIG. 5). The inhibition of angiogenesis formation from theinactivation of NF-kB by inhibiting a signal transduction was suggested.

Thus, the Psoraleae Semen extract, Siegesbeckiae Herba extract, andCorni Fructus extract of the present invention may be usefully used inangiogenesis inhibition and the prevention and treatment ofangiogenesis-related diseases.

The present invention also provides a composition for prevention andtreatment of angiogenesis-related diseases, containing one selected fromthe group consisting of Psoraleae Semen extract, Siegesbeckiae Herbaextract, and Corni Fructus extract as an active ingredient.

The present invention also provides a method for preventing or treatingangiogenesis-related diseases, including administering to an individualat least one selected from the group consisting of Psoraleae Semenextract, Siegesbeckiae Herba extract, and Corni Fructus extract.

The angiogenesis-related diseases include, but not limited to,cerebrovascular disease, cardiovascular disease, ocular disease orcancerous disease.

Folkman (PNAS 99:10730-10735, 2002) reported that a mouse lost weightwhen an angiogenesis inhibitor was administered to the mouse, andNishimura et al. (DIABETS, 56:1517-1526, 2007) revealed through laserconfocal microscopy technology that angiogenesis was directly associatedwith preadipocyte differentiation in obesity tissues by a 3D-imageanalysis technique. It was previously asserted that preadipocytes playeda leading role in angiogenesis by secreting various adipokines (VEGF,Leptin, TNF-a, and IL-6). However, Nishimura et al. disclosed that VEGFplayed a leading role in angiogenesis in adipose tissues. Therefore, atheory that obesity cell differentiation and proliferation may beprevented by inhibiting angiogenesis has been verified. Thus, an extracthaving angiogenesis inhibitory effects may be usefully used in treatmentof obesity.

Vascular maturation means that blood vessels which have been formedbecome functionally mature while another angiogenesis is inhibited, andcerebrovascular maturation is summarized as a process in which a bloodbrain barrier is developed and a selective permeability is induced.While the blood brain barrier is developed, the angiogenesis in thebrain is stopped and a capillary vessel is progressively differentiatedinto a blood brain barrier (Plate, K. H., J. Neuropathol. Exp. Neurol.,58, 313-320, 1999). Because extracts of the present invention allow fora functional maturation through the generation of a blood brain barrierby inhibiting angiogenesis, the extracts may be usefully used as agentsfor treatment and prevention of blood brain barrier dysfunction-relateddiseases. The angiogenesis-related cerebrovascular diseases include, butare not limited to, multiple sclerosis (Rosenberg G A, Neuroscientist,8(6):586-595, 2002), experimental allergic encephalomyelitis;Proescholdt M A et al., J Neuropathol Exp Neurol., 61(10):914-925,2002), bacterial meningitis, ischemia (Gashe Y et al., J Cere Blood FlowMetab., 21(12):1393-1400, 2001), brain edema (Dempsey R J et al.,Neurosurgery. 47(2):399-404; discussion 404-406, 2000), Alzheimer'sdisease (Banks W A et al., Peptides, 23(12):2223-2226, 2002), AcquiredImmune Deficiency Syndrome dementia complex, brain tumor, traumaticbrain injury; Esen F et al., J Neurosurg Anesthesiol., 15(2):119-125,2003), or hypertension (Kucuk M et al., Life Sci. 71(8):937-946 2002).

The cardiovascular diseases may be selected from the group consisting ofarteriosclerosis, vascular synechia, and scleredema (O'Brien K D et al.,Circulation 93(4):672-682, 1996), and the ocular diseases may beselected from the group consisting of keratoplastic angiogenesis,angiogenic glaucoma, macular degeneration, diabetic retinopathy,premature infant retinopathy, angiogenic glaucoma, angiogenesis mediatedcornea disease, macular degeneration, pterygium, retinal degeneration,retrolental fibroplasia, and granular conjunctivitis (Adamis A P et al.,Angiogenesis, 3:9-14, 1999).

The cancerous diseases may be selected from the group consisting ofastrocytoma, glioma, lung cancer, non-small cell lung carcinomas,hepatoma, colon carcinoma, bone cancer, pancreatic cancer, skin cancer,head or neck cancer, skin or intraocular melanoma, uterine cancer,ovarian cancer, rectal cancer, stomach cancer, cancer near the anus,colon carcinoma, breast cancer, fallopian tube carcinoma, endometrialcarcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma,Hodgkin's disease, esophageal cancer, small intestine cancer, endocrinegland cancer, thyroid cancer, parathyroid cancer, adrenal cancer, softtissue sarcoma, urethral cancer, penial cancer, prostatic cancer,bladder cancer, kidney or ureter cancer, renal cell carcinoma, pelviccarcinoma, central nervous system (CNS) tumor, primary CNS lymphoma,spinal cord tumor, and pituitary adenoma (Hanahan D et al., Cell,86:353-364, 1996), and the composition of the present invention may beapplied for the treatment of angiogenesis-related cancerous diseases andmetastasis.

The composition of the present invention may further include one or moreactive ingredients having the same or similar functions. The compositionof the present invention may also include one or more pharmaceuticallyacceptable carriers for administration. The composition of the presentinvention include about 0.0001% to about 10% by weight of the extractbased on the total weight, and preferably 0.001% to about 1% by weight.The pharmaceutically acceptable carrier may be prepared by mixing morethan one ingredient selected from the group consisting of saline,sterilized water, Ringer's solution, buffered saline, dextrose solution,maltodextrose solution, glycerol and ethanol. If necessary,anti-oxidative agent, buffer solution, bacteriostatic agent, and otherconventional additive may be added. Diluents, dispersing agents,surfactants, binders and lubricants may be additionally added to prepareinjectable solutions, pills, capsules, granules or tablets. Thecomposition of the present invention may further be preferably preparedfor each disease or according to ingredients by following suitablemethods in the art.

The composition of the present invention may be administered orally orparenterally (for example, intravenous, hypodermic, peritoneal or localapplication). The effective dosage of the composition may be determinedaccording to weight, age, gender, health condition, diet, administrationtime, administration method, excretion ratio and severity of a disease.The dosage of the present invention is about 0.01 to about 5,000 mg/kgper day, and preferably about 0.01 to about 10 mg/kg per day. Thecomposition is administered once a day or preferably a few times a day.

The present invention also provides a composition for inhibition ofobesity, containing Psoraleae Semen extract or Siegesbeckiae Herbaextract as an active ingredient.

The present invention also provides a method for inhibiting obesity,including administering to an individual an effective amount ofPsoraleae Semen extract or Siegesbeckiae Herba extract.

In a specific example of the present invention, the Psoraleae Semenextract or Siegesbeckiae Herba extract of the present inventionconcentration-dependently decreased adipose cells (See FIGS. 11 and 12).In addition, the Psoraleae Semen extract or Siegesbeckiae Herba extractof the present invention concentration-dependently decreased theexpression of PPARγ, a signal molecule which mediates the expression oflipogenesis, during an adipocyte differentiation process, and decreasedthe expressions of SREBP-1 and PPARγ, signal molecules which mediatelipogenesis even after a complete differentiation of adipocytes (SeeFIGS. 15 and 16). The Psoraleae Semen extract or Siegesbeckiae Herbaextract of the present invention decreased the weight in an animalexperiment when ingested with a high fat diet (See FIG. 17), andsignificantly decreased the levels of total serum cholesterol andneutral fat (See FIG. 4). Because Psoraleae Semen, Siegesbeckiae Herba,and Corni Fructus extracts of the present invention have effects oflipogenesis inhibition and adipogenesis inhibition, decrease weight, anddecrease the levels of total serum cholesterol and neutral fat, theextracts may be usefully used for inhibition of obesity.

The present invention also provides a health supplement food forpreventing or improving antiogenesis-related diseases, containing oneselected from the group consisting of Psoraleae Semen extract,Siegesbeckiae Herba extract, and Corni Fructus extract as an activeingredient.

Furthermore, the present invention provides a health supplement food forpreventing or improving obesity, containing Psoraleae Semen extract orSiegesbeckiae Herba extract as an active ingredient.

The Psoraleae Semen extract or Siegesbeckiae Herba extract of thepresent invention may be directly added, used with other foods or foodingredients, and suitably used according to conventional methods. Themixed amount of the active ingredients may be suitably determinedaccording to the purpose of use (prevention, health, or sanitation). Ingeneral, the Psoraleae Semen extract or Siegesbeckiae Herba extract ofthe present invention may be added in an amount of about 15 parts byweight or less with respect to raw material, and preferably about 10parts by weight or less. However, the amount may be below the aboverange in the case of a long-term administration for health orsanitation, or health management. Because there is no safety issuesregarding the extracts, the active ingredient may be used more than theabove ranges.

There are no specific limitations regarding the form and kind of thehealth supplement food. The health supplement food into which thePsoraleae Semen extract or Siegesbeckiae Herba extract may be added maybe in a form of tablet, capsule, powder, granule, liquid, and pill. Thehealth supplement food includes milk products including butter, yogurt,and cheese, dairy products including ice cream, bread, chocolate,candies, snacks, confections, ramyon (instant noodles), other noodles,gums, various soups, beverage, tea, drinks, alcoholic beverage, andvitamin complex, and all the health supplement foods in a conventionalsense.

The health supplement food of the present invention may contain variousflavors or natural carbohydrate as additional ingredients just likeconventional health supplement foods. The above natural carbohydrateincludes monosaccharide such as glucose and fructose, disaccharide suchas maltose and sucrose, polysaccharide such as dextrin and cyclodextrin,and sugar alcohol such as xylitol, sorbitol, and erythritol. Sweetenersmay include natural sweeteners such as thaumatin and stevia extract, andsynthetic sweeteners such as saccharin and aspartame. The content of thenatural carbohydrate may be generally about 0.01 g to about 0.04 g per100 Ml of the Psoraleae Semen extract or Siegesbeckiae Herba extract ofthe present invention, and preferably about 0.02 g to about 0.03 g.

In addition to the above, the health supplement food may contain variouskinds of nutrient, vitamin, electrolyte, flavoring agent, coloringagent, pectic acid and salt thereof, organic acid, preventive colloidalthickener, pH modifier, stabilizer, antiseptic, glycerin, alcohol,carbonation reagent used in the preparation of carbonated beverage, etc.In addition, the Psoraleae Semen extract or Siegesbeckiae Herba extractof the present invention may contain flesh of fruits to prepare naturalfruit juice, fruit juice beverage, and vegetable beverage. Thesecomponents may be used alone or in combination with other components.Although the content of these additives does not matter, the additivesmay be used in the range of about 0.01 to about 0.1 parts by weight per100 parts of the composition of the present invention.

Hereinafter, the present invention will be described in more detail withreference to the following examples and experimental examples.

However, the following examples and experimental examples are providedfor illustrative purposes only, and the scope of the present inventionshould not be limited thereto in any manner.

Example 1 Preparation of Extracts of Psoraleae Semen, SiegesbeckiaeHerba, and Corni Fructus

Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructus obtained from anherbal medicine shop (Daejeon, Korea) were confirmed through acomparative test of their external features by the Herbal QualityControl Team in Korea Institute of Oriental Medicine (KIOM) and thenused in an experiment. The Psoraleae Semen, Siegesbeckiae Herba, andCorni Fructus were washed, completely dried by a dryer, and cut intosmall pieces for use.

<1-1> Preparation of Water Extract

600 g of each chopped natural product was immersed in 5 times volume ofwater and hot water extracted for 5 hours. The extract was cooled down,subjected to a filtration process to obtain a supernatant, which wasconcentrated by an evaporator, and subjected to a freeze-drying toconcentrate the extract. 120 g (20%) of the extract was obtained.

<1-2> Preparation of Ethanol Extract

600 g of each chopped natural product was immersed in 5 times volume(3,000 Ml) of 70% ethanol, the mixture was refluxed for 5 hours whileheating, and an extract was obtained. The extraction process was 2 timesrepeated, and the extract was cooled down and subjected to a filtrationprocess to obtain a supernatant, which was later concentrated by anevaporator and subjected to a freeze-drying to concentrate the extract.90 g of the extract was obtained.

<1-3> Preparation of Methanol Extract

100% methanol was used in the same way as in Example 1-2 to obtain 100 gof an extract.

Experimental Example 1 Confirmation of Cell Adhesion Inhibitory Effects

<1-1> Confirmation of Cytotoxicity

HUVEC (Young Science, Korea) was seeded at 1×10⁴ cells/well in a 96-wellplate, incubated in EBM-2 medium (Cambrex, USA) supplemented with 2% FBSat 37° C. in a 5% CO₂-incubator, and further incubated until the maximumis reached. Subsequently, the extract in Example 1 was added at eachconcentration into the medium and incubated for 48 hours. Next, themedium in the culture solution was decanted, 1 Ml of 0.1 mM MTT wasadded, the mixture was incubated for another 4 hours, and the medium wasremoved. 200 μl of DMSO was aliquoted into each well and the absorbancewas measured at 570 nm.

As a result, as illustrated in FIG. 1, Psoraleae Semen, SiegesbeckiaeHerba, and Corni Fructus did not exhibit the cytotoxicity until 10 ppm,25 ppm, 25 ppm, respectively (FIG. 1).

<1-2> Confirmation of Cell Adhesion Factors

HUVEC (Young Science, Korea) was seeded at 5×10³ cells/well in a 96-wellplate, incubated in EBM-2 medium supplemented with 2% FBS at 37° C. in a5% CO₂-incubator, and further incubated until the maximum is reached.Subsequently, the extract in Example 1 was added at each concentrationinto the medium within the range of the cytotoxicity not observed andincubated for 20 hours. Next, the culture solution was washed with PBS,IL-1β (Endogen, USA) was added at 5 ng/Ml in order to induce celladhesion factors, and then the mixture was incubated for 6 hours. Themedium was removed, 1% paraformaldehyde was added, and a reaction wasperformed for 30 min to fix cells. After 30 min, the mixture was twicewashed with PBS+0.5% Tween 20 and blocked with 10% FBS for 1 hour.Monoclonal antibodies (VCAM-1, ICAM-1, and E-selectin: BD Bioscience,USA) were dissolved in PBS supplemented with 10% FBS at 2, 5, and 5μg/Ml respectively at 37° C. for 2 hours, and were washed with PBS+0.5%Tween 20. A secondary antibody (Donkey anti-mouse IgG-HRP: Bio-Rad, USA)was 1000-fold diluted in PBS supplemented with 10% FBS at roomtemperature for 1 hour, a coloring agent (Western Blotting LuminolReagent: Bio-Rad, USA) was added into the dilution, reaction wasperformed for 1 hour, and then the absorbance was measured at 405 nm ona plate reader (UVM 340, ASYS, Australia).

As a result, as illustrated in Tables 1 to 3, it was observed that theethanol extract among the water, ethanol, and methanol extractssufficiently inhibited the cell adhesion promoting factors VCAM-1,ICAM-1, and E-selectin. Thus, a subsequent experiment was performed withan ethanol extract (See Tables 1 to 3).

TABLE 1 Inhibition rates of VCAM-1 by Psoraleae Semen, SiegesbeckiaeHerba, and Corni Fructus Concentration Water Ethanol Methanol Sample(ppm) Extract Extract Extract Negative control — — — — (physiologicalsaline solution) Psoraleae Semen 1  98.2% 100.0% 95.0% SiegesbeckiaeHerba 5 110.0%   114% 102.0%  Corni Fructus 5 100.0% 111.8% 98.7%

TABLE 2 Inhibition rates of ICAM-1 by Psoraleae Semen, SiegesbeckiaeHerba, and Corni Fructus Concentration Water Ethanol Methanol Sample(ppm) Extract Extract Extract Negative control — — — — (physiologicalsaline solution) Psoraleae Semen 1 98.0% 100.0%    95% SiegesbeckiaeHerba 5  128%  136%  110% Corni Fructus 5 65.0% 66.7% 63.2%

TABLE 3 Inhibition rates of E-selectin by Psoraleae Semen, SiegesbeckiaeHerba, and Corni Fructus Concentration  Water Ethanol Methanol Sample(ppm) Extract Extract Extract Negative control — — — — (physiologicalsaline solution) Psoraleae Semen 1 120% 128.0% 105% Siegesbeckiae Herba5 115% 120.0% 100% Corni Fructus 5 97.5%  100.7% 96.0% 

As a result, aS illustrated in FIGS. 2 to 4, each extract of PsoraleaeSemen, Siegesbeckiae Herba, and Corni Fructus concentration-dependentlyinhibited the expressions of cell adhesion promoting factors VCAM-1,ICAM-1, and E-selectin. Specifically, Psoraleae Semen, SiegesbeckiaeHerba, and Corni Fructus exhibited significantly lower inhibitoryeffects in the case of VCAM-1 at 1.0 ppm, 5.0 ppm, and 5.0 ppm,respectively compared to a control group, and it was confirmed that theinhibitory strengths were in the order of Psoraleae Semen (0.5 ppm,100%)>Siegesbeckiae Herba (5.0 ppm, 114%)>Corni Fructus (5.0 ppm,111.8%). It was observed in the case of ICAM-1 that the inhibitorystrengths were in the order of Psoraleae Semen (0.5 ppm,100%)>Siegesbeckiae Herba (5.0 ppm, 138%)>Corni Fructus (5.0 ppm,66.7%), while it was observed in the case of E-selectin that theinhibitory strengths were in the order of Psoraleae Semen (1.0 ppm,128%)>Siegesbeckiae Herba (5.0 ppm, 120%)>Corni Fructus (5.0 ppm,100.7%). In conclusion, it was confirmed that the inhibitory strengthswere in the order of Psoraleae Semen (5.0 ppm)>Siegesbeckiae Herba (0.5ppm)>Corni Fructus (5.0 ppm) (FIGS. 2 to 4).

Experimental Example 2 Confirmation of Inhibitory Effects ofAngiogenesis

150 μl of matrigel was added into a 24-well plate (Becton DickinsonLabware, USA) to coat the surface and allowed to stand at 37° C. for 1hour for solidification. Cells (2.5×10⁴ cells/well) separated by addingtrypsin-EDTA into the well plate were aliquoted, the extracts in Example1 were administered at each concentration into the aliquots, and themixtures were incubated under 37° C. and 5% CO₂ conditions for 4 hours.Subsequently, once tube networks were formed, five samples were randomlyselected and photographed by a digital camera (Coolpix; Nikon, Japan) toobtain images, and tube lengths were measured by an NIH image program.

As a result, as illustrated in FIG. 5, it was confirmed that theangiogenesis inhibitory effects by Psoraleae Semen, Siegesbeckiae Herba,and Corni Fructus extracts were excellent compared to those by EGCG as anegative control (FIG. 5).

Experimental Example 3 Confirmation of Angiogenesis InhibitoryMechanisms

In order to understand the angiogenesis inhibitory mechanisms byPsoraleae Semen, Siegesbeckiae Herba, and Corni Fructus extracts, thepresent inventors examined signal transduction molecules β-catenin andvascular epithelium-cadherin (VE-cadherin); and the subgroup Akt(Protein kinase B) to study the actions of the extracts.

<3-1> Extract Processing and VEGF Stimulation

Specifically, 1×10⁵ cells of HUVEC were inoculated into a 100 mmpetri-dish and incubated. When the maximum was reached, the extracts inExample 1 were processed at each concentration, incubated for 24 hours,and washed with serum-free EBM-2 medium. The processed group wasincubated overnight without addition of serum, VEGF (BD Science, USA)was added at a final concentration of 50 ng/Ml and a reaction wasperformed at 37° C. for 30 min.

<3-2> Protein Isolation and Immunoprecipitation

In order to prevent phosphorylation, vanadate (100 μM) and hydrogenperoxide (200 μM) were added for 7 min prior to the lysis of cells.Subsequently, cells were washed with PBS+0.1 mM Na₃VO₄, which wasimmediately cooled down, and a cooled lysis buffer [150 mM NaCl, 10 mMTris-HCl (pH 7.4), 1% Triton X-100, 1 mM vanadate, 1 mM EDTA, 1 mM FGTA,0.2 mM PMSF, 0.5% NP-40] was added to perform a reaction at 4° C. for 20min. Next, cells were harvested with a stripper and introduced into amicroependorf tube, followed by centrifugation at 4° C. and 14,000 rpmfor 10 min. The concentrations of proteins in the supernatant werequantified by using a BCA protein test kit (Pierce, USA), andimmunoprecipitations were performed by using an anti-VE-Cadherinantibody (BD Bioscience, USA). 50 μg of the proteins and 10 μl of theantibody were added into 500 μl of immunoprecipitation buffer(supplemented with 0.2 mM sodium vanadate), and then H₂O was added tomake 1 Ml as the total volume. After the working solution was vortexed,a reaction was performed at 4° C. for 12 hours and 20 μl of proteinA/G-agarose (Amersham Science, Sweden) was added. After another reactionat 4° C. for 30 min, a centrifugation was performed at 4° C. and 14,000rpm for 3 min to remove the supernatant. The solution was washed with500 μl of a washing buffer and then subjected to a centrifugation toremove the supernatant.

<3-3> Western Blotting

50 μl of 4× Laemmli sample loading buffer was added into a purifiedprotein and boiled at 95° C. for 5 min. The sample was subjected toelectrophoresis with 7% SDS-PAGE gel and transferred to 0.2 μmnitrocellulose membrane (Bio-Rad Lab. USA). The protein transferred intothe membrane was identified with a Ponceau-S (Markham, Canada) reagent,blocked with 5%-non-fat dry milk (NFDM), and each treated with primaryantibodies anti-β-catenin, anti-Akt, anti-VE-cadherin, and anti-β-actin(BD Bioscience, USA), respectively. A secondary antibody donkeyanti-goat IgG-HRP was diluted at a ratio of 1:7000 for reaction and wasexposed to a film by using ECL (Santa Cruz Biotechnology, USA) as afluorescent material.

As a result, as illustrated in FIGS. 6 and 7, it was confirmed that thedetection signals of the antibodies decreased as the concentrations ofthe extracts (FIGS. 6 and 7). From these, the extractsconcentration-dependently inhibited the expression of a signaltransduction molecule, and the inhibition of the signal transductionprevented the signal transduction molecule from being transferred to theNF-kB (FIG. 9). Thus, it was proposed that the inactivation of the NF-kBinhibit angiogenesis.

Experimental Example 4 Confirmation of In Vivo Angiogenesis InhibitoryEffects of Natural Products by a Chorioallantoic Membrane (CAM) Test

After a fertilized egg was grown in a 37° C. incubator in which thehumidity of 70% or more was maintained for 3 days and perforated, 4 Mlof albumin was withdrawn by a syringe. When the fertilized egg became anembryo at day 4, a round window was drilled on an upper part of the airsac in the fertilized egg and a transparent tape was put on the part. Inorder to observe the angiogenesis inhibitory effects, 200 μl ofPsoraleae Semen, Siegesbeckiae Herba, and Corni Fructus extracts at 10,25, and 25 ppm, respectively were applied to 13 mm-diameter thermatoxcoverslips and dried. When the embryo was grown for 4.5 days, thetransparent tape was detached, these coverslips were placed on thesurface of chorioallantoic membrane (CAM) of a developing embryo, andthen the window was blocked with the transparent tape. A thermatoxcoverslip was only attached on the chorioallantoic membrane (CAM) ofchick embryo as a control group. After this was incubated in anincubator for 2 days, 10% fat emulsion was injected with a syringe intothe chorioallantoic membrane (CAM) and the angiogenesis process wasobserved. 60 or more chorioallantoic membranes of chick embryo were usedfor each experimental group and photographed by a digital camera toobtain their images.

As a result, as illustrated in FIG. 10, it was confirmed that theextract of the present invention significantly inhibited in-vivoangiogenesis (FIG. 10).

Experimental Example 5 Confirmation of Adipocyte Inhibitory Effects

3T3-L1 preadipocytes were incubated in a DMEM/10% FBS medium until thebottom of the flask was covered with the cells, and further incubatedfor another 2 days. An induction medium (MDI, 0.5 mM3-isobutyl-1-methylxanthine, 0.5 μM dexamethasone, 10 μg/Ml insulin) wasplaced into the culture for incubation to induce the differentiation ofthe preadipocytes into the adipocytes. The medium was exchanged with anormal DMEM/10% FBS medium on day 3, and Psoraleae Semen, SiegesbeckiaeHerba, and Corni Fructus extracts were processed at each concentrationfor 7 to 10 days while the media were exchanged once in two days.

In order to analyze the differentiation into the adipocytes, theincubated cells were fixed with formalin. After the formalin wasremoved, the cells were washed with 60% isopropanol and dried untilwhite kernels appeared. After the cells were stained with oil-red and aworking solution, the staining solution was removed and washed fourtimes with distilled water. After a staining solution on non-fatportions was washed with 100% isopropanol, a stained intracellular fatwas observed under microscope.

As a result, as illustrated in FIGS. 11 to 13, it was observed thatPsoraleae Semen, Siegesbeckiae Herba, and Corni Fructus extractssignificantly decreased adipocytes in a concentration-dependent manner(FIGS. 11 to 13).

Experimental Example 6 Lipogenesis and Adipogenesis Inhibitory Effects

In order to confirm the degrees of adipogenesis and lipogenesisinhibition by Psoraleae Semen, Siegesbeckiae Herba, and Corni Fructusextracts at the protein level, the present inventors performed a Westernblot. In order to confirm the inhibitions of lipogenesis andadipogenesis by two signal molecules associated with the lipogenesissignal pathway, the present inventors used SREBP-1 (SC-367) andPPARγ(SC-7273) antibodies to perform a Western blot on a group in whicha differentiation material and a sample were simultaneously introducedand a group in which a sample was introduced after a differentiation wasinduced, respectively (FIG. 14).

<6-1> 3T3-L1 Cell Differentiation and Sample Treatment

After 3T3-L1 cells were pre-incubated to fill a T-75 flask, the cellswere trypsinized to aliquot 1×10⁵ cells on each p100 dish. DMEMcontaining 10% BS was added into the dishes to make 10 Ml as a totalvolume and the incubation was performed at 37° C. under 5% CO₂conditions

When cells were fully incubated, they were incubated for two more days.A first group was simultaneously treated with a differentiation inducingmaterial MDI (0.5 mM 3-isobutyl-1-methyl-xanthine, 1 μM dexamethasone,and 10 μg/Ml insulin), Psoraleae Semen, Siegesbeckiae Herba, and CorniFructus extracts. The medium was removed after two days and changed intoa medium containing DMEM/10% FBS, insulin, and sample. The medium waschanged into a medium containing only DMEM/10% FBS, and sample afteranother two days. After a differentiation of 3T3-L1 preadipocytes intoadipocytes was completed in a group in which a sample was introducedafter a differentiation was induced, Psoraleae Semen, SiegesbeckiaeHerba, and Corni Fructus extracts were treated a total three times ateach concentration (1, 5, and 10 ppm) at an interval of two days.

<6-2> Protein Isolation and Quantification

In order to obtain proteins from 3T3-L1 cells, a medium was removed, thecells were twice with cold PBS (supplemented with 1 mM vanadate), RIPAlysis buffer (SC-24948) (50 mM Tris-Cl pH 7.4, 1% NP-40, 0.25% sodiumdeoxycholate, 0.1% SDS, 150 mM NaCl, 1 mM EDTA) was introduced at 600μl/dish, and a shake incubation was performed at 4° C. for 10 min. Afterthe culture was subjected to a centrifugation at 4° C. and 12,000 rpmfor 20 min, only a transparent supernatant was collected, transferred toan ependorf tube, and stored at −80° C. The concentration of the proteinwas measured using a BCA reagent in the same manner as in HUVECs. Theconcentration was calculated according to the absorbance and treatedwith 1× Laemmli Sample Buffer at each concentration. A hot block wasused to boil samples at 95° C. for 10 min and the samples were stored at−80° C.

<6-3> SDS-PAGE, Membrane Transfer and Film Phenomena

The measured protein was used to perform an electrophoresis and membranetransfer. In order to confirm the specific expression pattern of a3T3-L1 adipocyte protein, two antibodies SREBP-1 and PPARγ, signalmolecules associated with adipogenesis, were reacted at room temperatureunder none-fat-dry-milk (NFDM) conditions for 1 hour, diluted at a ratioof 1:1000 with 1% NFDM, and then stood still at 4° C. overnight. Afterthe reaction was completed, the antibodies were washed four times with1×TBST and a reaction was performed with a secondary antibody at roomtemperature for 1 hour. After the antibodies were washed four times with1×TEST and a reaction was performed with a coloring agent for 5 min, afilm was developed in a dark room.

As a result, when each of Psoraleae Semen, Siegesbeckiae Herba, andCorni Fructus extracts was treated simultaneously with MDI as adifferentiation inducing material, it was confirmed that the expressionlevel of PPARγ, a signal molecule which mediates the expression oflipogenesis, decreased as the concentrations of the extracts increase asillustrated in FIG. 15. In particular, excellent lipogenesis inhibitoryeffects were observed at 10 ppm of Psoraleae Semen, Siegesbeckiae Herba,and Corni Fructus extracts (FIG. 15).

When 3T3-L1 cells were completely differentiated and then PsoraleaeSemen, Siegesbeckiae Herba, and Corni Fructus extracts were treated ateach concentration, it was confirmed that as illustrated in FIG. 16, theexpression levels of SREBP-1 and PPARγ, signal molecules which mediatethe expression of lipogenesis, decreased as the concentrations of theextracts increased (FIG. 16).

Therefore, it was confirmed that Psoraleae Semen, Siegesbeckiae Herba,and Corni Fructus extracts have lipogenesis inhibitory effects.

Experimental Example 7 Confirmation of Weight Loss Effects

<7-1> Measurement of Weight Changes

After Sprague-Dawley rats were fed a high fat diet for 16 days, 1 Ml ofPsoraleae Semen extract at 2 mg/200 g (10 mg/kg), Siegesbeckiae Herbaextract at 5 mg/200 g (10 mg/kg), and Corni Fructus at 5 mg/200 g (10mg/kg) were orally administered for 15 days while they were fed a highfat diet to observe the weight changes. A control group was fed only ahigh fat diet without administering the extract.

As a result, as illustrated in FIG. 17, when a high fat diet wasadministered in combination with Psoraleae Semen, Siegesbeckiae Herba,and Corni Fructus extracts for 15 days, weight losses were observed by27.1%, 34.1%, and 23.5%, respectively (FIG. 17).

<7-2> Blood Analysis

Blood was collected from the rats in <7-1> and analyzed.

As a result, as illustrated in the following Table 4, the totalcholesterol levels were significantly reduced according to theadministration of samples to male rats, compared to a control group. Inparticular, the neutral fat levels were significantly reduced except forin xenical as a positive control, and the HDL-cholesterols fell within anormal range (40 to 50 mg/dl). The total cholesterol levels in femalerats were not significantly reduced compared to a control group.However, the neutral fat levels were significantly reduced compared tothe level in xenical as a positive control

TABLE 4 Ingredient AST ALT T-CHO TG HDL-C Sample (IU/L) (IU/L) (mg/dl)(mg/dl) (mg/dl) Mcon 524 98 174 124 28 MP 200 50 125 55 51 MH 216 56 12866 50 MS 130 42 132 52 52 MX 116 38 116 150 50 Fcon 298 58 110 80 36 FP154 44 106 58 36 FH 184 42 100 54 38 FS 218 88 110 56 34 FX 160 40 10378 36 Mcon: male control; MP: male pagoji (Psoraleae semen); MH: maleheuichum (Siegesbekiae herba); MS: male sansuyu (Corni fructus); MX:male xenical; AST(U/L): GOT; ALT(U/L): GPT; T-CHO: Cholesterol (mg/dL):200 mg/dl or less; TG: Triglyceride (mg/dL): 150 mg/dl or less; andHDL-C: HDL Cholesterol: 40-50 mg/dl or more.

Therefore, it was found that Psoraleae Semen, Siegesbeckiae Herba, andCorni Fructus induced a weight loss, accompanied by a decrease inneutral fat.

In summary, because the extracts of the present invention exhibitedanti-angiogenesis effects, induced a weight loss in a short timeaccording to the effects, and decreased the levels of the totalcholesterol and neutral fat, each extract of the Psoraleae Semen,Siegesbeckiae Herba, and Corni Fructus of the present invention may beusefully used as an anti-obesity agent.

Preparation Example 1 Preparation of Pharmaceutical Formulations

1. Preparation of Powder

Extract of the present invention 2 g Lactose 1 g

The components were mixed and filled in an airtight sac to prepare apowder agent.

2. Preparation of Tablet

Extract of the present invention 100 mg Corn starch 100 mg Lactose 100mg Magnesium stearate  2 mg

The components were mixed and a tableting was performed according to aconventional tablet preparation method to prepare a capsule.

3. Preparation of Capsule

Extract of the present invention 100 mg Corn starch 100 mg Lactose 100mg Magnesium stearate  2 mg

The components were mixed and filled into a gelatin capsule according toa conventional capsule preparation method to prepare a capsule.

4. Preparation of Pill

Extract of the present invention   1 g Lactose 1.5 g Glycerin   1 gXylitol 0.5 g

The components were mixed and prepared such that 4 g per pill wasobtained according to a conventional method.

5. Preparation of Granule

Extract of the present invention 150 mg Soybean extract  50 mg Glucose200 mg Starch 600 mg

The components were mixed, 100 mg of 30% ethanol was added, and themixture was dried at 60° C. to form a granule, which was filled in asac.

Preparation Example 2 Preparation of Food

Foods including an extract of the present invention were prepared in thefollowing manner.

1. Preparation of Flour Food

Food for health improvement was prepared by adding the extract of thepresent invention by 0.5% to 5% by weight into wheat flour, and thenbread, cakes, cookies, crackers and noodles were prepared by using themixture.

2. Preparation of Dairy Products

Various dairy products such as butter and ice cream were prepared byadding the extract of the present invention by 5% to 10% by weight intomilk and using the milk.

Preparation Example 3 Preparation of Beverages

Extract of the present invention 1000 mg Citric acid 1000 mgOligosaccharide 100 g Mumefural concentrate 2 g Taurine 1 g Total byaddition of purified water 900 Ml

According to a conventional heal beverage preparation method, thecomponents are mixed and heated with stirring at 85° C. for about 1hour. A resulting solution is filtered and decanted into a sterilized 2l container with sealed and sterilized. The container is stored in arefrigerator for further use as health supplement foods of the presentinvention.

Although the compositions were prepared by mixing components relativelyappropriate for fancy drinks in a preferred example, they may bemodified considering demand class, demand country, purpose of use, localand national preference, etc.

Because the Psoraleae Semen extract, Siegesbeckiae Herba extract, andCorni Fructus extract of the present invention exhibit inhibitoryeffects of angiogenesis and inhibitory effects of obesity, they may beusefully used as active ingredients for a composition for prevention andtreatment of angiogenesis-related diseases or obesity.

The Psoraleae Semen extract, Siegesbeckiae Herba extract, and CorniFructus extract of the present invention may be usefully used indevelopment of pharmaceutical composition for prevention or treatment ofangiogenesis-related diseases or obesity, or of health supplement foodfor prevention or improvement of angiogenesis-related diseases.

Although the preferred embodiments of the present invention have beendisclosed for illustrative purposes, those skilled in the art willappreciate that various modifications, additions and substitutions arepossible, without departing from the scope and spirit of the inventionas disclosed in the accompanying claims.

What is claimed is:
 1. A method for treating obesity, comprisingadministering to an individual in need thereof, a composition consistingessentially of an effective amount of a 70% ethanol extract ofSiegesbeckiae Herba.
 2. The method of claim 1, wherein the extract ofSiegesbeckiae Herba inhibits angiogenesis.
 3. The method of claim 1,wherein the composition is in a form selected from the group consistingof injectable solution, pill, capsule, granule and tablet.
 4. The methodof claim 1, wherein administering comprises administering about 0.01 toabout 5,000 mg/kg per day to said individual.
 5. The method of claim 4,wherein administering comprises administering about 0.01 to about 10mg/kg per day to said individual.
 6. The method of claim 4, wherein thecomposition is in a form selected from the group consisting of butter,yogurt, cheese, ice cream, bread, chocolate, candy, a confection,noodles, gum, soup, a beverage, tea, a drink, an alcoholic beverage anda vitamin complex.
 7. The method of claim 4, wherein the compositionfurther comprises a food additive selected from the group consisting ofa nutrient, a vitamin, an electrolyte, a flavoring agent, a coloringagent, pectic acid or a salt thereof, an organic acid, a colloidalthickener, a pH modifier, a stabilizer, an antiseptic, glycerin, alcoholor carbonation.
 8. The method of claim 4, wherein the composition is inthe form of a fruit or vegetable juice.
 9. The method of claim 1,wherein the composition further comprises a pharmaceutically acceptablecarrier.
 10. The method of claim 9, wherein the pharmaceuticallyacceptable carrier is selected from the group consisting of saline,sterilized water, Ringer's solution, buffered saline, dextrose solution,maltodextrose solution, glycerol, ethanol, an antioxidant, buffersolution, bacteriostatic agent, diluent, dispersing agent, surfactant,binder, lubricant and combinations thereof.
 11. The method of claim 9,wherein the pharmaceutically acceptable carrier is present in thecomposition from about 0.0001% to about 10% by weight of thecomposition.
 12. The method of claim 11, wherein the pharmaceuticallyacceptable carrier is present in the composition from 0.001% to about 1%by weight of the composition.